Zhang Yifan, Yuan Jin, Zhang Yizhu, Chen Ye, Cao Jiaoxian, An Rui, Wang Xinhong
School of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; School of Pharmacy, Shanghai Institute of Health Science, Shanghai, 201318, China.
Biomed Chromatogr. 2015 Apr;29(4):485-95. doi: 10.1002/bmc.3300. Epub 2014 Aug 18.
A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of 12 constituents of Gegen Qinlian Decoction (GQD), namely puerarin, daidzein, baicalin, wogonoside, wogonin, liquiritin, liquiritigenin, berberine, jatrorrhizine, palmatine, coptisine and glycyrrhetic acid, in rat plasma. The plasma samples were spiked with the internal standard (IS) carbamazepine acidified with HCl and extracted by liquid-liquid extraction with ethyl acetate. Chromatographic separation was achieved on a Shiseido Capcell PAK C18 column utilizing a gradient elution profile and a mobile phase consisting of (A) 0.1% formic acid in water and (B) acetonitrile. Detection was performed in the multiple reaction monitoring mode using electrospray ionization in the positive ion mode at a flow rate of 0.3 mL/min and a run time of 8 min. All of the calibration curves gave good linearity (r > 0.9930) over the concentration range from 0.6-360 to 16.2-9720 ng/mL for all components. The intra- and inter-day precisions were <15.0% in terms of the relative standard deviation, and the accuracies were within ±13.7% in terms of the relative error. The method was successfully applied to investigate the pharmacokinetics of the major active compounds of Gegen Qinlian Decoction after its oral administration to rats.
建立了一种液相色谱-串联质谱(LC-MS/MS)法并进行了验证,用于同时测定大鼠血浆中葛根芩连汤(GQD)的12种成分,即葛根素、大豆苷元、黄芩苷、汉黄芩苷、汉黄芩素、甘草苷、甘草素、小檗碱、药根碱、巴马汀、黄连碱和甘草次酸。血浆样品加入内标(IS)卡马西平,用盐酸酸化,然后用乙酸乙酯液-液萃取。采用梯度洗脱,流动相由(A)0.1%甲酸水溶液和(B)乙腈组成,在资生堂Capcell PAK C18柱上实现色谱分离。采用电喷雾电离正离子模式,在多反应监测模式下进行检测,流速为0.3 mL/min,运行时间为8 min。所有成分在0.6 - 360至16.2 - 9720 ng/mL浓度范围内的校准曲线均具有良好的线性(r > 0.9930)。日内和日间精密度的相对标准偏差<15.0%,准确度的相对误差在±13.7%以内。该方法成功应用于大鼠口服葛根芩连汤后主要活性成分的药代动力学研究。